Physicochemical, structural and biological characterisation of poly(3-hydroxyoctanoate) supplemented with diclofenac acid conjugates - Harnessing the potential in the construction of materials for skin regeneration processes.

This study involved creating oligomeric conjugates of 3-hydroxy fatty acids and diclofenac, named Dic-oligo(3HAs). Advanced NMR techniques confirmed no free diclofenac in the mix. We tested diclofenac release under conditions resembling healthy and chronic wound skin. These oligomers were used to make P(3HO) blends, forming patches for drug delivery. Their preparation used the solvent casting/porogen leaching (SCPL) method. The patches' properties like porosity, roughness, and wettability were thoroughly analysed. Antimicrobial assays showed that Dic-oligo(3HAs) exhibited antimicrobial activity against reference (S. aureus, S. epidermis, S. faecalis) and clinical (Staphylococcus spp.) strains. Human keratinocytes (HaCaT) cell line tests, as per ISO 10993-5, showed no toxicity. A clear link between material roughness and HaCaT cell adhesion was found. Deep cell infiltration was verified using DAPI and phalloidin staining, observed under confocal microscopy. SEM also confirmed HaCaT cell growth on these scaffolds. The strong adhesion and proliferation of HaCaT cells on these materials indicate their potential as wound dressing layers. Additionally, the successful diclofenac release tests point to their applicability in treating both normal and chronic wounds.

Life cycle assessment of experimental Al-ion batteries for energy storage applications.

In this work, the analysis of environmental performance and its coherence with circular economy priorities of different variants of Al-ion battery construction was performed. Al-ion-based batteries can be considered as one of the future alternatives for currently used Li-ion-based cells when the shortage of lithium or cobalt becomes a challenge. All tested batteries were constructed with Al anodes, polypropylene foil separator, polyvinylidene fluoride + N-Methyl-2-pyrrolidone (PVDF+NMP) binder, Al collector and laminated Al foil pouch cell. WO3, Norit and carbon from potato starch (CPS) were used as a cathode material. Saturated solutions of AlCl3 dissolved in diethylene glycol dimethyl ether (DEG) and deep eutectic solvents (DES) originating from bacterial polymer polyhydroxyalkanoate were used as electrolytes. The ReCiPe impact assessment method was used in this analysis. The indicator in this study was ReCiPe Endpoint (H) V1.07 referring to Europe. SimaPro 9.4 software with Ecoinvent 3.8 inventory database were used for all calculations. The analysis included experimental production and assembly of batteries and their end-of-life processing. Based on the performed analysis it was found that the overall weighted impact of each single construction variant of an Al-ion battery is dominated by the use of electricity, no matter which variant is considered since it is related to the electricity mix in Poland and its high dependence on fossil fuels. Overall environmental impact is the smallest for CPS DEG battery, while Norit DEG and CPS DEG variants have slightly higher impacts. The share of end-of-life processing in overall environmental impacts of all analysed variants was found low compared to the Li-ion batteries. This observation indicates the Al-ion batteries as a promising direction of alternative electrochemical devices for energy storage systems while end-of-life processing and circular solution are concerned.

Upcycling Waste Streams from a Biorefinery Process-A Case Study on Cadmium and Lead Biosorption by Two Types of Biopolymer Post-Extraction Biomass.

This study investigated the possibility of using the spent kind of biomass of Pseudomonas putida CA-3 and Zobelella denitrificans MW1 obtained after the pilot-scale production of polyhydroxyalkanoates (PHAs) as a biosorbent for the bioremediation of aqueous solutions containing toxic cadmium and lead ions. The material was characterized by means of scanning electron microscopy, Fourier-transformed infrared spectroscopy, nuclear magnetic resonance spectroscopy and amino acid profiling. To check the sorption capacity of spent biomass against Pb and Cd ions, equilibrium studies were performed. To learn about the nature of the sorption process, kinetic modelling was carried out and the obtained results showed that the adsorption process is best described by the pseudo-second-order kinetic model (PSO), which suggests that the sorption process is connected with the chemical bonding of the ions on the sorbent surface. Information provided by the amino acid profile made it possible to predict the adsorption mechanism and FTIR analysis proved the participation of different chemical groups in the removal process. According to the equilibrium studies, the best-fitted isotherm was the Freundlich model for all used materials and metal ions considering the correlation coefficient. Summarizing the results, the spent biomass after the PHA production is an effective biosorbent and can be reused for heavy metal bioremediation.

Biological and physiochemical studies of electrospun polylactid/polyhydroxyoctanoate PLA/P(3HO) scaffolds for tissue engineering applications.

Polyhydroxyoctanoate, as a biocompatible and biodegradable biopolymer, represents an ideal candidate for biomedical applications. However, physical properties make it unsuitable for electrospinning, currently the most widely used technique for fabrication of fibrous scaffolds. To overcome this, it was blended with polylactic acid and polymer blend fibrous biomaterials were produced by electrospinning. The obtained PLA/PHO fibers were cylindrical, smaller in size, more hydrophilic and had a higher degree of biopolymer crystallinity and more favorable mechanical properties in comparison to the pure PLA sample. Cytotoxicity evaluation with human lung fibroblasts (MRC5 cells) combined with confocal microscopy were used to visualize mouse embryonic fibroblasts (MEF 3T3 cell line) migration and distribution showed that PLA/PHO samples support exceptional cell adhesion and viability, indicating excellent biocompatibility. The obtained results suggest that PLA/PHO fibrous biomaterials can be potentially used as biocompatible, biomimetic scaffolds for tissue engineering applications.

Physicochemical studies of novel sugar fatty acid esters based on (R)-3-hydroxylated acids derived from bacterial polyhydroxyalkanoates and their potential environmental impact.

Sugar fatty acids esters are popular compounds widely used in both the nutritional, cosmetic and pharmaceutical industries due to their amphiphilic structure and consequent ability to reduce the surface tension of solutions. Furthermore, an important aspect in the implementation of any additives and formulations is their environmental impact. The properties of the esters depend on the type of sugar used and the hydrophobic component. In this work, selected physicochemical properties of new sugar esters based on lactose, glucose and galactose and hydroxy acids derived from bacterial polyhydroxyalkanoates are shown for the first time. Values for critical aggregation concentration, surface activity and pH make it possible that these esters could compete with other commercially used esters of similar chemical structure. The investigated compounds showed moderate emulsion stabilization abilities presented on the example of water-oil systems containing squalene and body oil. Their potential environmental impact appears to be low, as the esters are not toxic to Caenorhabditis elegans even at concentrations much higher than the critical aggregation concentration.

The Influence of Novel, Biocompatible, and Bioresorbable Poly(3-hydroxyoctanoate) Dressings on Wound Healing in Mice.

The human body's natural protective barrier, the skin, is exposed daily to minor or major mechanical trauma, which can compromise its integrity. Therefore, the search for new dressing materials that can offer new functionalisation is fully justified. In this work, the development of two new types of dressings based on poly(3-hydroxyoctanoate) (P(3HO)) is presented. One of the groups was supplemented with conjugates of an anti-inflammatory substance (diclofenac) that was covalently linked to oligomers of hydroxycarboxylic acids (Oli-dicP(3HO)). The novel dressings were prepared using the solvent casting/particulate leaching technique. To our knowledge, this is the first paper in which P(3HO)-based dressings were used in mice wound treatment. The results of our research confirm that dressings based on P(3HO) are safe, do not induce an inflammatory response, reduce the expression of pro-inflammatory cytokines, provide adequate wound moisture, support angiogenesis, and, thanks to their hydrophobic characteristics, provide an ideal protective barrier. Newly designed dressings containing Oli-dicP(3HO) can promote tissue regeneration by partially reducing the inflammation at the injury site. To conclude, the presented materials might be potential candidates as excellent dressings for wound treatment.

Study on ßTCP/P(3HB) Scaffolds-Physicochemical Properties and Biological Performance in Low Oxygen Concentration.

The search for new materials for bone regenerative purposes is still ongoing. Therefore, we present a series of newly constructed composites based on ß tricalcium phosphate (ßTCP) and poly(3-hydroxybutyrate) bacteria-derived biopolymer (P(3HB)) in the form of 3D scaffolds with different pore sizes. To improve the polymer attachment to the ßTCP surface, the etching of ceramic sinters, using citric acid, was applied. As expected, pre-treatment led to the increase in surface roughness and the creation of micropores facilitating polymer adhesion. In this way, the durability and compressive strength of the ceramic-polymer scaffolds were enhanced. It was confirmed that P(3HB) degrades to 3-hydroxybutyric acid, which broadens applications of developed materials in bone tissue engineering as this compound can potentially nourish surrounding tissues and reduce osteoporosis. Moreover, to the best of our knowledge, it is one of the first studies where the impact of ßTCP/P(3HB) scaffolds on mesenchymal stem cells (MSCs), cultured in lowered (5%) oxygen concentration, was assessed. It was decided to use a 5% oxygen concentration in the culture to mimic the conditions that would be found in damaged bone in a living organism during regeneration. Scaffolds enabled cell migration and sufficient flow of the culture medium, ensuring high cell viability. Furthermore, in composites with etched ßTCP, the MSCs adhesion was facilitated by hydrophilic ceramic protrusions which reduced hydrophobicity. The developed materials are potential candidates for bone tissue regeneration. Nevertheless, to confirm this hypothesis, in vivo studies should be performed.

A polyhydroxyalkanoates bioprocess improvement case study based on four fed-batch feeding strategies.

The modelling and optimization of a process for the production of the medium chain length polyhydroxyalkanoate (mcl-PHA) by the bacterium Pseudomonas putida KT2440 when fed a synthetic fatty acid mixture (SFAM) was investigated. Four novel feeding strategies were developed and tested using a constructed model and the optimum one implemented in further experiments. This strategy yielded a cell dry weight of 70.6 g l-1 in 25 h containing 38% PHA using SFAM at 5 l scale. A phosphate starvation strategy was implemented to improve PHA content, and this yielded 94.1 g l-1 in 25 h containing 56% PHA using SFAM at 5 l scale. The process was successfully operated at 20 l resulting in a cell dry weight of 91.2 g l-1 containing 65% PHA at the end of a 25-h incubation.

COVID-19 lockdown shows how much natural mountain regions are affected by heavy tourism.

Mountain areas in Poland are among the most frequented tourist destinations and such intensive tourism negatively affects the natural environment. The COVID-19 pandemic and the resulting lockdown restricted travel for a few months in 2020, providing a unique opportunity to observe the studied mountain environment without the impact of typical tourist traffic. This study is based on the determination of antibiotic content, hydrochemical parameters, enumeration of culturable bacterial water quality indicators, antimicrobial susceptibility tests together with extended spectrum beta-lactamase (ESBL) gene detection in waterborne E. coli and NGS-based bacterial community composition at six sites along the Bialka river valley (one of the most popular touristic regions in Poland) in three periods: in summer and winter tourist seasons and during the COVID-19 lockdown. The results of individual measurements showed decreased numbers of bacterial indicators of water contamination (e.g. numbers of E. coli dropped from 99 × 104 CFU/100 ml to 12 CFU/100 ml at the most contaminated site) and the share of antimicrobial resistant E. coli (total resistance dropped from 21% in summer to 9% during lockdown, share of multidrug resistant strains from 100 to 44%, and ESBL from 20% in summer to none during lockdown). Antibiotic concentrations were the highest during lockdown. The use of multivariate analysis (principal component analysis - PCA and heatmaps) revealed a clear pattern of tourism-related anthropogenic pressure on the water environment and positive impact of COVID-19 lockdown on water quality. PCA distinguished three major factors determining water quality: F1 shows strong effect of anthropogenic pressure; F2 describes the lockdown-related quality restoration processes; F3 is semi-natural and describes the differences between the most pristine and most anthropogenically-impacted waters.

Polyhydroxyalkanoates, bacterially synthesized polymers, as a source of chemical compounds for the synthesis of advanced materials and bioactive molecules.

Research into polyhydroxyalkanoates (PHAs) is growing exponentially. These bacterially derived polyesters offer a spectrum of possible applications, such as in manufacturing of daily-use objects, production of medical devices and implantable objects, or as synthons in chemical and pharmaceutical industries. Thanks to their broad physicochemical features, PHAs can be seen as polymers of the future, which can replace traditional petrochemical equivalents. As they are synthesized by bacteria through fermentation processes, these polyesters can be obtained from virtually any carbon source in a sustainable manner. Characterized by biodegradability and biocompatibility, they are used in many industries, ranging from production of everyday objects to medical applications. Furthermore, as they are built from bioactive monomers, namely (R)-3-hydroxyacids, they provide a platform for the synthesis of advanced chemical compounds. In this mini review, the reader will be acquainted with recent studies conducted at the Jerzy Haber Institute of Catalysis and Surface Chemistry of the Polish Academy of Sciences in collaboration with other groups that have contributed to the development of PHA-based medical materials, bioactive molecules and novel green solvents derived from PHA monomers.Key points• Polyhydroxyalkanoates are emerging polymers for biomedical applications• Polyhydroxyalkanoates can be modified easily to provide novel materials• (R)-3-Hydroxyacids are good synthons for bioactive substances and green solvents.

Robust process for high yield conversion of non-degradable polyethylene to a biodegradable plastic using a chemo-biotechnological approach.

In this study, the optimisation of a process for producing medium-chain-length polyhydroxyalkanoate (mcl-PHA) by Pseudomonas putida KT2440 when fed with a polyethene (PE)-derived fatty acid mixture was investigated. The PE was pyrolysed to produce a hydrocarbon wax that was subsequently oxidised to produce a mixture of fatty acids, purified, and used as a PHA substrate for the growth and selection of microorganisms. Based on the shaken flask screening, a production strain, i.e., Pseudomonas putida KT2440, was selected for conducting bioreactor studies. Feeding PE-derived fatty acids in a 20-L setup resulted in high mcl-PHA yields (83.0 g L-1 CDW with 65% PHA in 25 h). Furthermore, life-cycle assessment (LCA) was conducted to determine the environmental advantages of the proposed process and its impacts compared to those of other technologies for treating PE-derived waste streams. We conclude that processing waste PE into PHA, rather than incineration, produces biodegradable material while also reducing the additional emissions that arise from traditional PE waste treatment processes, such as incineration to gain energy.

Silver Decorated ßTCP-Poly(3hydroxybutyrate) Scaffolds for Bone Tissue Engineering.

Implantations in orthopedics are associated with a high risk of bacterial infections in the surgery area. Therefore, biomaterials containing antibacterial agents, such as antibiotics, bactericidal ions or nanoparticles have been intensively investigated. In this work, silver decorated ß tricalcium phosphate (ßTCP)-based porous scaffolds were obtained and coated with a biopolymer-poly(3-hydroxybutyrate)-P(3HB). To the best of our knowledge, studies using silver-doped ßTCP and P(3HB), as a component in ceramic-polymer scaffolds for bone tissue regeneration, have not yet been reported. Obtained materials were investigated by high-temperature X-ray diffraction, X-ray fluorescence, scanning electron microscopy with energy dispersive spectroscopy, hydrostatic weighing, compression tests and ultrahigh-pressure liquid chromatography with mass spectrometry (UHPLC-MS) measurements. The influence of sintering temperature (1150, 1200 °C) on the scaffolds' physicochemical properties (phase and chemical composition, microstructure, porosity, compressive strength) was evaluated. Materials covered with P(3HB) possessed higher compressive strength (3.8 ± 0.6 MPa) and surgical maneuverability, sufficient to withstand the implantation procedures. Furthermore, during the hydrolytic degradation of the composite material not only pure (R)-3-hydroxybutyric acid but also its oligomers were released which may nourish surrounding tissues. Thus, obtained scaffolds were found to be promising bone substitutes for use in non-load bearing applications.

Vimentin Cytoskeleton Architecture Analysis on Polylactide and Polyhydroxyoctanoate Substrates for Cell Culturing.

Polylactide (PLA), widely used in bioengineering and medicine, gained popularity due to its biocompatibility and biodegradability. Natural origin and eco-friendly background encourage the search of novel materials with such features, such as polyhydroxyoctanoate (P(3HO)), a polyester of bacterial origin. Physicochemical features of both P(3HO) and PLA have an impact on cellular response 32, i.e., adhesion, migration, and cell morphology, based on the signaling and changes in the architecture of the three cytoskeletal networks: microfilaments (F-actin), microtubules, and intermediate filaments (IF). To investigate the role of IF in the cellular response to the substrate, we focused on vimentin intermediate filaments (VIFs), present in mouse embryonic fibroblast cells (MEF). VIFs maintain cell integrity and protect it from external mechanical stress, and also take part in the transmission of signals from the exterior of the cell to its inner organelles, which is under constant investigation. Physiochemical properties of a substrate have an impact on cells' morphology, and thus on cytoskeleton network signaling and assembly. In this work, we show how PLA and P(3HO) crystallinity and hydrophilicity influence VIFs, and we identify that two different types of vimentin cytoskeleton architecture: network "classic" and "nutshell-like" are expressed by MEFs in different numbers of cells depending on substrate features.

Polyhydroxyalkanoate/Antifungal Polyene Formulations with Monomeric Hydroxyalkanoic Acids for Improved Antifungal Efficiency.

Novel biodegradable and biocompatible formulations of "old" but "gold" drugs such as nystatin (Nys) and amphotericin B (AmB) were made using a biopolymer as a matrix. Medium chain length polyhydroxyalkanoates (mcl-PHA) were used to formulate both polyenes (Nys and AmB) in the form of films (~50 µm). Thermal properties and stability of the materials were not significantly altered by the incorporation of polyenes in mcl-PHA, but polyene containing materials were more hydrophobic. These formulations were tested in vitro against a panel of pathogenic fungi and for antibiofilm properties. The films containing 0.1 to 2 weight % polyenes showed good activity and sustained polyene release for up to 4 days. A PHA monomer, namely 3-hydroxydecanoic acid (C10-OH), was added to the films to achieve an enhanced synergistic effect with polyenes against fungal growth. Mcl-PHA based polyene formulations showed excellent growth inhibitory activity against both Candida yeasts (C. albicans ATCC 1023, C. albicans SC5314 (ATCC MYA-2876), C. parapsilosis ATCC 22019) and filamentous fungi (Aspergillus fumigatus ATCC 13073; Trichophyton mentagrophytes ATCC 9533, Microsporum gypseum ATCC 24102). All antifungal PHA film preparations prevented the formation of a C. albicans biofilm, while they were not efficient in eradication of mature biofilms, rendering them suitable for the transdermal application or as coatings of implants.

In Search of Effective Anticancer Agents-Novel Sugar Esters Based on Polyhydroxyalkanoate Monomers.

Cancer is one of the deadliest illness globally. Searching for new solutions in cancer treatments is essential because commonly used mixed, targeted and personalized therapies are sometimes not sufficient or are too expensive for common patients. Sugar fatty acid esters (SFAEs) are already well-known as promising candidates for an alternative medical tool. The manuscript brings the reader closer to methods of obtaining various SFAEs using combined biological, chemical and enzymatic methods. It presents how modification of SFAE's hydrophobic chains can influence their cytotoxicity against human skin melanoma and prostate cancer cell lines. The compound's cytotoxicity was determined by an MTT assay, which followed an assessment of SFAEs' potential metastatic properties in concentrations below IC50 values. Despite relatively high IC50 values (63.3-1737.6 µM) of the newly synthesized SFAE, they can compete with other sugar esters already described in the literature. The chosen bioactives caused low polymerization of microtubules and the depolymerization of actin filaments in nontoxic levels, which suggest an apoptotic rather than metastatic process. Altogether, cancer cells showed no propensity for metastasis after treating them with SFAE. They confirmed that lactose-based compounds seem the most promising surfactants among tested sugar esters. This manuscript creates a benchmark for creation of novel anticancer agents based on 3-hydroxylated fatty acids of bacterial origin.

A study on the structure, mechanism, and biochemistry of kanamycin B dioxygenase (KanJ)-an enzyme with a broad range of substrates.

Kanamycin A is an aminoglycoside antibiotic isolated from Streptomyces kanamyceticus and used against a wide spectrum of bacteria, including Mycobacterium tuberculosis. Biosynthesis of kanamycin involves an oxidative deamination step catalyzed by kanamycin B dioxygenase (KanJ), thereby the C2' position of kanamycin B is transformed into a keto group upon release of ammonia. Here, we present for the first time, structural models of KanJ with several ligands, which along with the results of ITC binding assays and HPLC activity tests explain substrate specificity of the enzyme. The large size of the binding pocket suggests that KanJ can accept a broad range of substrates, which was confirmed by activity tests. Specificity of the enzyme with respect to its substrate is determined by the hydrogen bond interactions between the methylamino group of the antibiotic and highly conserved Asp134 and Cys150 as well as between hydroxyl groups of the substrate and Asn120 and Gln80. Upon antibiotic binding, the C terminus loop is significantly rearranged and Gln80 and Asn120, which are directly involved in substrate recognition, change their conformations. Based on reaction energy profiles obtained by density functional theory (DFT) simulations, we propose a mechanism of ketone formation involving the reactive FeIV  = O and proceeding either via OH rebound, which yields a hemiaminal intermediate or by abstraction of two hydrogen atoms, which leads to an imine species. At acidic pH, the latter involves a lower barrier than the OH rebound, whereas at basic pH, the barrier leading to an imine vanishes completely. DATABASES: Structural data are available in PDB database under the accession numbers: 6S0R, 6S0T, 6S0U, 6S0W, 6S0V, 6S0S. Diffraction images are available at the Integrated Resource for Reproducibility in Macromolecular Crystallography at http://proteindiffraction.org under DOIs: 10.18430/m36s0t, 10.18430/m36s0u, 10.18430/m36s0r, 10.18430/m36s0s, 10.18430/m36s0v, 10.18430/m36s0w. A data set collection of computational results is available in the Mendeley Data database under DOI: 10.17632/sbyzssjmp3.1 and in the ioChem-BD database under DOI: 10.19061/iochem-bd-4-18.

Physicochemical and Biological Characterisation of Diclofenac Oligomeric Poly(3-hydroxyoctanoate) Hybrids as ß-TCP Ceramics Modifiers for Bone Tissue Regeneration.

Nowadays, regenerative medicine faces a major challenge in providing new, functional materials that will meet the characteristics desired to replenish and grow new tissue. Therefore, this study presents new ceramic-polymer composites in which the matrix consists of tricalcium phosphates covered with blends containing a chemically bounded diclofenac with the biocompatible polymer-poly(3-hydroxyoctanoate), P(3HO). Modification of P(3HO) oligomers was confirmed by NMR, IR and XPS. Moreover, obtained oligomers and their blends were subjected to an in-depth characterisation using GPC, TGA, DSC and AFM. Furthermore, we demonstrate that the hydrophobicity and surface free energy values of blends decreased with the amount of diclofenac modified oligomers. Subsequently, the designed composites were used as a substrate for growth of the pre-osteoblast cell line (MC3T3-E1). An in vitro biocompatibility study showed that the composite with the lowest concentration of the proposed drug is within the range assumed to be non-toxic (viability above 70%). Cell proliferation was visualised using the SEM method, whereas the observation of cell penetration into the scaffold was carried out by confocal microscopy. Thus, it can be an ideal new functional bone tissue substitute, allowing not only the regeneration and restoration of the defect but also inhibiting the development of chronic inflammation.

What Has Been Trending in the Research of Polyhydroxyalkanoates? A Systematic Review.

Over the past decades, enormous progress has been achieved with regard to research on environmentally friendly polymers. One of the most prominent families of such biopolymers are bacterially synthesized polyhydroxyalkanoates (PHAs) that have been known since the 1920s. However, only as recent as the 1990s have extensive studies sprung out exponentially in this matter. Since then, different areas of exploration of these intriguing materials have been uncovered. However, no systematic review of undertaken efforts has been conducted so far. Therefore, we have performed an unbiased search of up-to-date literature to reveal trending topics in the research of PHAs over the past three decades by data mining of 2,227 publications. This allowed us to identify eight past and current trends in this area. Our study provides a comprehensive review of these trends and speculates where PHA research is heading.

Vimentin Association with Nuclear Grooves in Normal MEF 3T3 Cells.

Vimentin, an intermediate filament protein present in leukocytes, blood vessel endothelial cells, and multiple mesenchymal cells, such as mouse embryonic fibroblasts (MEF 3T3), is crucial for various cellular processes, as well as for maintaining the integrity and durability (stability) of the cell cytoskeleton. Vimentin intermediate filaments (VIFs) adhere tightly to the nucleus and spread to the lamellipodium and tail of the cell, serving as a connector between the nucleus, and the cell's edges, especially in terms of transferring mechanical signals throughout the cell. How these signals are transmitted exactly remains under investigation. In the presented work, we propose that vimentin is involved in that transition by influencing the shape of the nucleus through the formation of nuclear blebs and grooves, as demonstrated by microscopic observations of healthy MEF (3T3) cells. Grooved, or "coffee beans" nuclei, have, to date, been noticed in several healthy cells; however, these structures are especially frequent in cancer cells-they serve as a significant marker for recognition of multiple cancers. We observed 288 MEF3T3 cells cultured on polyhydroxyoctanoate (PHO), polylactide (PLA), and glass, and we identified grooves, coaligned with vimentin fibers in the nuclei of 47% of cells cultured on PHO, 50% of cells on glass, and 59% of cells growing on PLA. We also observed nuclear blebs and associated their occurrence with the type of substrate used for cell culture. We propose that the higher rate of blebs in the nuclei of cells, cultured on PLA, is related to the microenvironmental features of the substrate, pH in particular.

How sustainable are biopolymers? Findings from a life cycle assessment of polyhydroxyalkanoate production from rapeseed-oil derivatives.

The main purpose of the article was to compare different scenarios of biopolymer production and their impacts on the environment using Life Cycle Assessment. Three alternative polyhydroxyalkanoates (PHA: amorphous PHA and poly(3-hydroxybutyrate), P(3HB)) production scenarios were considered to assess its environmental impact: Scenario A - Production of mcl-PHA/P(3HB) from crude vegetable oil; Scenario B - Production of P(3HB) with biodiesel by-product; Scenario C - Production of mcl-PHA/P(3HB) from used vegetable oil. Subject to the scenario considered, it was shown that the environmental efficiency of PHA production is highly dependent on carbon sources used, and it is strongly supporting production of mcl-PHA instead of P(3HB). As LCA study shows, due to low yield of P(3HB) in comparison to mcl-PHA production in considered processes, all the P(3HB) production scenarios have higher impacts than the production of mcl-PHA. Production processes based on bacterial fermentation had its impacts related mostly to the raw materials used and to its separation phase. Additionally, using secondary materials instead of raw ones, namely used oil instead of virgin oil, gives significant improvement with regard to environmental impact. The resource efficiency is also the identified as the key factor with sensitivity analysis that indicates the possible increase of biopolymer yield as the most beneficial factor. Biobased polymers have big environmental potential but still need significant improvement with regard to their manufacturing processes in order to become more economically benign. Preferably production of these microbial polymers should be integrated into biorefinery blocks, where such waste stream arises (e.g. biodiesel production plant).